A simple method was developed to isolate enzymatic and metabolic active fraction of cerebral microvessels from nonvascular brain tissue which showed a saturable uptake of 2-deoxy-D-(3H) glucose. The Km for 3H 2-deoxy-D-glucose was 0.1 mM and the uptake was inhibited by the non-metabolizable 3-0-methyl-D-glucose and the inhibitors of carrier transport - phlorizin and phloretin.